Hyperpolarization of 13C-labeled energy substrates enables the noninvasive detection and mapping of metabolic activity, in vivo, with magnetic resonance spectroscopy (MRS). Therefore, hyperpolarization and 13C MRS can potentially become a powerful tool to study the physiology of organs such as the heart, through the quantification of kinetic patterns under both normal and pathological conditions. In this study we assessed myocardial uptake and metabolism of hyperpolarized [1-13 C]pyruvate in anesthetized pigs. Pyruvate metabolism was studied at baseline and during dobutamine-induced stimulation. We applied a numerical approach for spectral analysis and kinetic fitting (LSFIT/KIMOfit), making a comparison with a well-known jMRUI/AMARES analysis and γ-variate function, and we estimated the apparent conversion rate of hyperpolarized [1-13 C]pyruvate into its downstream metabolites [1-13C]lactate, [1-13C]alanine and [13C]bicarbonate in a 3 T MR scanner. We detected an increase in the apparent kinetic constants (kPX) for bicarbonate and lactate of two-fold during dobutamine infusion. These data correlate with the double product (rate-pressure product), an indirect parameter of cardiac oxygen consumption: we observed an increase in value by 46 ± 11% during inotropic stress. The proposed approach might be applied to future studies in models of cardiac disease and/or for the assessment of the pharmacokinetic properties of suitable 13C-enriched tracers for MRS. Copyright © 2012 John Wiley & Sons, Ltd.